Mouse Anti-Cyclic N-oxide 5 Recombinant Antibody (clone 4C5) (CAT#: PABC-030)

Figure 1 Electrophoretic analysis of mAb 4C5, followed by immunoblotting with an anti-mouse kappa chain.

Intact IgG1 immunoglobulin produced by the 2D10 hybridoma cells serves as positive control.

SIDERA, Katerina, et al. The 4C5 cell-impermeable anti-HSP90 antibody with anti-cancer activity, is composed of a single light chain dimer. PloS one, 2011, 6.9.

Figure 2 Electrophoretic mobility and specificity of antibodies.

Reverse immunoprecipitation experiments in MDA-MB-453 celllysates using mAb 4C5, rec-4C5 and ch-4C5, followed by western blotwith anti-HSP90a. In all cases a single immunoreactive band is observed indicating that both recombinant antibodies recognize HSP90.

SIDERA, Katerina, et al. The 4C5 cell-impermeable anti-HSP90 antibody with anti-cancer activity, is composed of a single light chain dimer. PloS one, 2011, 6.9.

Figure 3 Immunofluoresence detection of intracellular HSP90 in fixed MDA-MB-453 cells, permeabilized with 0.1% Triton X-100.

Similarly to the murine antibody, rec-4C5 and ch-4C5 recognize the intracellular pool of HSP90. Scale bar: 20 mm

SIDERA, Katerina, et al. The 4C5 cell-impermeable anti-HSP90 antibody with anti-cancer activity, is composed of a single light chain dimer. PloS one, 2011, 6.9.

Figure 4 Visualization of dead cells using trypan blue dye.

In ch-4C5 treated cultures, the cell death incidence is similar to that observed in the control cultures. In contrast, in the cultures treated with the anti-HSP90a antibody, a much greater number of cells are stained with Trypan blue, indicating an increased incidence of cell death. Scale bar, 30 mm.

SIDERA, Katerina, et al. The 4C5 cell-impermeable anti-HSP90 antibody with anti-cancer activity, is composed of a single light chain dimer. PloS one, 2011, 6.9.

Figure 5 ch-4C5 inhibis cancer cell invasion in vitro.

Control and ch-4C5 treated cells were fixed permeabilized and stained with fluorescently labelled phalloidin. Scale bar: 40 mm

SIDERA, Katerina, et al. The 4C5 cell-impermeable anti-HSP90 antibody with anti-cancer activity, is composed of a single light chain dimer. PloS one, 2011, 6.9.

Figure 6 ch-4C5 inhibits the metastatic deposition of MDA-MB-453 cancer cells into the lungs of SCID mice.

Representative cryosections of the lungs of control and ch-4C5 treated mice. The arrows show MDA-MB-453 cells stained with DiO present in the lung tissue. A significant decrease in the deposition of cancer cells was observed in the lungs of ch-4C5 treated mice. Scale bar: 100 mm

SIDERA, Katerina, et al. The 4C5 cell-impermeable anti-HSP90 antibody with anti-cancer activity, is composed of a single light chain dimer. PloS one, 2011, 6.9.